• Mass Spectrometry Letters
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• 제4권1호
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• pp.1-9
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• 2013

Fungal biotechnology has been well established in food and healthcare sector, and now being explored for lignocellulosic biorefinery due to their great potential to produce a wide array of extracellular enzymes for nutrient recycling. Due to global warming, environmental pollution, green house gases emission and depleting fossil fuel, fungal enzymes for lignocellulosic biomass refinery become a major focus for utilizing renewal bioresources. Proteomic technologies tender better biological understanding and exposition of cellular mechanism of cell or microbes under particular physiological condition and are very useful in characterizing fungal secretome. Hence, in addition to traditional colorimetric enzyme assay, mass-spectrometry-based quantification methods for profiling lignocellulolytic enzymes have gained increasing popularity over the past five years. Majority of these methods include two dimensional gel electrophoresis coupled to mass spectrometry, differential stable isotope labeling and label free quantitation. Therefore, in this review, we reviewed more commonly used different proteomic techniques for profiling fungal secretome with a major focus on two dimensional gel electrophoresis, liquid chromatography-based quantitative mass spectrometry for global protein identification and quantification. We also discussed weaknesses and strengths of these methodologies for comprehensive identification and quantification of extracellular proteome.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1895-1902
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• 2008

Adipose tissue is an important endocrine organ involved in the control of whole body energy homeostasis and insulin sensitivity. Considering the increased incidence of obesity and obesity-related disorders, including diabetes, it is important to understand thoroughly the process of adipocyte differentiation and its control. Therefore, we performed a differential proteome mapping strategy using two-dimensional gel electrophoresis combined with peptide mass fingerprinting to identify intracellular proteins that are differentially expressed during adipose conversion of 3T3-L1 pre-adipocytes in response to an adipogenic cocktail. In the current study, we identified 46 differentially expressed proteins, 6 of which have not been addressed previously in 3T3-L1 cell differentiation. Notably, we found that phosphoribosyl pyrophosphate synthetase (PRPS), a regulator of cell proliferation, was preferentially expressed in pre-adipocytes than in fully differentiated adipocytes. In conclusion, our results provide valuable information for further understanding of the adipogenic process.

• Animal cells and systems
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• 제4권2호
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• pp.145-150
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• 2000

Retinoic acid (RA) evokes pattern duplication in the regenerating salamander limb. Interestingly, it also enhances dedifferentiation in the regenerate by the morphological, histological and biochemical criteria. To examine whether there is any correlation between the RA-evoked pattern duplication and de novo protein synthetic profile in the regenerating salamander limb, especially during dedifferentiation, we analyzed stage-specific protein synthesis pattern in the normal and RA-treated regenerating limbs by metabolic labeling followed by two-dimensional gel electrophoresis. In the regenerating limbs without RA treatment, a few hundred kinds of proteins were found to be synthesized at the stage of wound healing and the total number of protein synthesized increased greatly as regeneration proceeded. The same trend was also observed in the RA-treated regenerating limbs. Interestingly, some protein spots were noted to be either newly synthesized or highly expressed by the RA treatment especially at the stage of dedifferentiation. The results shows that the enhancement of dedifferentiation state after the RA treatment correlates well with the protein synthesis profile, and suggest that those proteins are important for the RA-evoked pattern duplication in the regenerating limbs of salamander.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1221-1229
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• 2019

Mycobacterium tuberculosis, a causative pathogen of tuberculosis (TB), still threatens human health worldwide. To find a novel drug to eradicate this pathogen, we tested taurine-5-bromosalicylaldehyde Schiff base (TBSSB) as an innovative anti-mycobacterial drug using Mycobacterium smegmatis as a surrogate model for M. tuberculosis. We investigated the antimicrobial activity of TBSSB against M. smegmatis by plotting growth curves, examined the effect of TBSSB on biofilm formation, observed morphological changes by scanning electron microscopy and transmission electron microscopy, and detected differentially expressed proteins using two-dimensional gel electrophoresis coupled with mass spectrometry. TBSSB inhibited mycobacterial growth and biofilm formation, altered cell ultrastructure and intracellular content, and inhibited cell division. Furthermore, M. smegmatis adapted itself to TBSSB inhibition by regulating the metabolic pathways and enzymatic activities of the identified proteins. NDMA-dependent methanol dehydrogenase, NAD(P)H nitroreductase, and amidohydrolase AmiB1 appear to be pivotal factors to regulate the M. smegmatis survival under TBSSB. Our dataset reinforced the idea that Schiff base-taurine compounds have the potential to be developed as novel anti-mycobacterial drugs.

• 한국작물학회지
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• 제50권spc1호
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• pp.4-7
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• 2005

미강으로부터 정제된 tocotrienol은 DPPH를 기질로 확인한 결과 매우 뛰어난 항산화력을 가지는 것으로 판명되었다. 또한 정상세포와 암세포를 배양하면서 tocotrienol을 처리하고 세포내의 항산화에 가장 큰 역할을 하는 superoxide dismutase와 glutathione peroxidase 활성을 측정한 결과 두 효소 모두 tocotrienol에 의하여 활성이 증가되는 것을 볼 수 있었으며, 전체적으로 암세포에서 GPX가 SOD보다 더 민감하게 증가함을 알 수 있었다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.147.2-147.2
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• 2003

A fast and matrix-assisted laser desorption/ionization-mass spectrometry compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, Zincon and Ethyl Violet to form an ion-pair complex. It is safe to use since the methanol used previously in staining solution was replaced with ethanol, which is not toxic. The protocol including fixing, staining and quick washing steps can be completed in 1 to 1.5 h depending upon gel thickness. (omitted)

• 한국해양바이오학회지
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• 제2권1호
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• pp.68-72
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• 2007

넙치의 혈청 단백질체를 연구하기 위해 본 연구에서는 이차원전기영동 분석 시의 기본조건을 확립하였다. 넙치의 혈청단백질을 이차원전기영동 법으로 분석하기 위해서는 TCA 침전을 이용하여 혈청 내에 고농도로 존재하는 이온물질을 제거하는 전처리과 정이 반드시 요구되었다. 또한 분석 단백질의 농도는 $30{\mu}g$이 적당하였다. 넙치 10개체의 혈청단백질을 이용하여 총 51회의 이차원전기영동을 수행한 결과 1,8207개의 단백질로 이루어진 넙치 혈청단백질 표준 지도를 작성할 수 있었다.

• 한국동물학회지
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• 제35권1호
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• pp.17-22
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• 1992

The three yolk polypeptides have been isolated and partially characterized. Their molecular weights of YPI, YP2, and YP3 were 48, 000, 47, 000, and 46, 000, respectivelv, as judged by SDS-polyacrvlamide gel electrophoresis. They have different digestion products upon in situ peptide mapping by limited proteolysis. Two-dimensional gel electrophoresis showed that their isoelectric points were heterogeneous from 5.92 to 6.54. And thew showed three different antigen-antibody reactions when each polvpeptides is reacted with antisera made to a mixture of all of three. These data reported here indicate that the yolk proteins are consisted of distinctive polypeptides in Drosophlla sp. (robusta species group).

• 정보처리학회논문지B
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• 제12B권3호
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• pp.239-246
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• 2005

생물학자가 단백질을 검색하고 분석하기 위해서는 2차원 젤 전기영동(2DGE : Two Dimensional Gel Electrophoresis) 실험을 해야 한다. 실험 결과는 2차원 영상이 생성된다. 2차원 영상에서 단백질 반점의 패턴 분석을 위해 2차원 젤 영상에 펼쳐진 단백질 반점들을 영상처리를 통해 분할하고, 대조 그룹의 단백질 패턴과 비교분석을 통해 밝히고자하는 단백질 반점을 찾아내야 한다. 단백질 반점을 분할하는 알고리즘에 있어서 기존에는 가우시안 함수를 적용하였지만, 최근 들어 형태학 분리개념에 의한 Watersheds 영역기반 분할(Watersheds region-based segmentation) 알고리즘을 활용하고 있다. 그러나 Watersheds 영역기반 분할 알고리즘은 크기가 큰 영상에서 원하는 영역을 신속하게 분할한다는 장점이 있지만, 영상 화소의 그레이 값이 연속적인 경우 실제 반점의 개수 에 비해 과다분할(over-segmentation)되거나 과소분할(under-segmentation)의 문제점을 안고 있다. 이는 마커(marker) 포인트의 설정에 의해 어느 정도 해결할 수 있지만 병합(merge)과 분할(split) 과정을 반복해야 한다. 본 논문은 Watersheds 기반 계층적 이진화 기법을 적용하여 마커 드리븐 Watersheds 영상분할의 문제점을 해결하고자 한다.

• The Korean Journal of Physiology and Pharmacology
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• 제8권4호
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• pp.207-211
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• 2004

The purpose of the present study was to evaluate the expression of cardiac marker protein in rabbit cardiac tissue that was exposed to ischemic preconditioning (IPC), or ischemiareperfusion injury (IR) using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). We compared 2DE gels of control (uninjured) cardiac tissue with those of IPC and IR cardiac tissue. Expression of one protein was detected in IR heart tissue, however the protein was not detected in the samples of control and IPC tissue. To further characterize the detected protein molecule, the protein in the 2D gel was isolated and subjected to trypsin digestion, followed by MALDI-MS. The protein was identified as myoglobin, which was confirmed also by Western blot analysis. These results are consistent with previous studies of cardiac markers in ischemic hearts, indicating myoglobin as a suitable marker of myocardial injury. In addition, the present use of multiple techniques indicates that proteomic analysis is an appropriate means to identify cardiac markers in studies of IPC and IR.